VEGF secretion in MDA-MB-231 cell line - (Dec/16/2008 )

Hello experts,

Can someone tell me if this breast cancer cell line (MDA-MB-231) secretes VEGF if grown in a flask (in-vitro) naturally? or do i need to induce it with something?

Thanks

-Pronana-

Hi,

Count the cell Plate it (40,000/well in 96well or 24well plate)Treat compound at desired conc (triplicate). Keep untreated controlsincubate it for 24,48 and 72hcollect the supernatantsSubject it for ELISA ( R&D system or any other)

Sure you will detect VEGF.

Best regards

-kamxeno-

QUOTE (kamxeno @ Dec 16 2008, 08:02 PM)

Hi,

Count the cell Plate it (40,000/well in 96well or 24well plate)Treat compound at desired conc (triplicate). Keep untreated controlsincubate it for 24,48 and 72hcollect the supernatantsSubject it for ELISA ( R&D system or any other)

Sure you will detect VEGF.

Best regards

Hi there,I usually plate my cells with DMEM supplemented with 10% FCS. Will this effect the reading? is it necessary for me to use serum-free DMEM media when i incubate the cells for 24, 48, and 72h? Thanks for your help. very much grateful

-Pronana-

QUOTE (Pronana @ Dec 17 2008, 08:06 PM)

QUOTE (kamxeno @ Dec 16 2008, 08:02 PM)

Hi,

Count the cell Plate it (40,000/well in 96well or 24well plate)Treat compound at desired conc (triplicate). Keep untreated controlsincubate it for 24,48 and 72hcollect the supernatantsSubject it for ELISA ( R&D system or any other)

Sure you will detect VEGF.

Best regards

Hi there,I usually plate my cells with DMEM supplemented with 10% FCS. Will this effect the reading? is it necessary for me to use serum-free DMEM media when i incubate the cells for 24, 48, and 72h? Thanks for your help. very much grateful

Hello,Greetings.... DMEM with 10% FCS is fine. If you want to do deep research on VEGF, use with out serum, 5% and 10% under different duration with various cell line etc etc .....

But now you are going to screen your extract on VEGF secertion. So, at this junction DMEM with 10%FCS, three time point and 5diff conc of extarct is fine to show inhibitory potential.